Minimal disease monitoring by QRT-PCR: Guidelines for identification and systematic validation of molecular markers prior to evaluation in prospective clinical trials

V. F. Viprey, M. A. Lastowska, M. V. Corrias, K. Swerts, M. S. Jackson, S. A. Burchill

Research output: Contribution to journalArticle

Abstract

Real-time RT-PCR (QRT-PCR) is a sensitive method for the detection of minimal disease (MD) and may improve monitoring of disease status and stratification of patients for therapy. Where tumour-specific mRNAs have not been identified, the selection of which target(s) is(are) optimal for the detection of MD remains a challenge. This reflects the heterogeneity of tumour cells, the stability of mRNAs and low-level of transcription in cells of the normal haemopoietic compartments. The aim of this study was to establish for the first time guidelines for the systematic prioritization of potential markers of MD detected by QRT-PCR prior to evaluation in multicentre prospective clinical outcome studies. We combined microarray analysis, ESTs gene expression profiles, improved probe-sets sequence annotation, and previously described standard operating procedures for QRT-PCR analysis to identify and prioritize potential markers of MD. Using this methodology, we identified 49 potential markers of MD in neuroblastoma (NB), of which 11 were associated with neuronal function. We found that, in addition to TH, Phox2B and DCX mRNA may be useful targets for the detection of MD in children with NB. This same strategy could be exploited to select MD markers of other solid tumours from the large number of potential targets identified by microarray gene expression profiles.

Original languageEnglish
Pages (from-to)245-252
Number of pages8
JournalJournal of Pathology
Volume216
Issue number2
DOIs
Publication statusPublished - Oct 2008

Keywords

  • Bone marrow
  • Microarrays
  • Minimal residual disease
  • Neuroblastoma
  • Peripheral blood
  • Real-time RT-PCR

ASJC Scopus subject areas

  • Pathology and Forensic Medicine

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